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user manualchromatography software

um 03-0014-92

Getting StartedWith

Getting Started with UNICORN 5.003-0014-92 Edition AB2004-02

Office addresses:

Amersham Biosciences ABSE-751 84 UppsalaSweden

Amersham Biosciences UK LimitedAmersham PlaceLittle ChalfontBuckinghamshireEngland HP7 9NA

Amersham Biosciences Corp.800 Centennial AvenueP.O. Box 1327Piscataway NJ 08855USA

Amersham Biosciences Europe GmbHMunzinger Strasse 9D-79111 FreiburgGermany

Amersham Biosciences K.K.Sanken Building3-25-1 Hyakunincho, Shinjuku-kuTokyo 169-0073Japan

Amersham Biosciences China Limited13/F., Tower IEver Gain Plaza88 Container Port RoadKwai Chung, New TerritoriesHong Kong

www.amershambiosciences.com

Trademarks:

UNICORN, Drop Design, BioProcess, ÄKTA and ÄKTAexplorer are trademarks of Amersham Biosciences Limited.Amersham and Amersham Biosciences are trademarks of Amersham plc.Microsoft and Windows are trademarks of the Microsoft Corporation in the United States and/or other countries.

Terms and Condition of Sale:

Unless otherwise agreed in writing, all goods and services are sold subject to the terms and conditions of the company within the Amersham Biosciences group which supplies them. A copy of these terms and conditions is available on request. Any use of this software is subject to Amersham Biosciences Standard Software End-User License Agreement.

© Copyright Amersham Biosciences AB 2004 - All rights reserved.

Table of Contents

1. Introducing UNICORN..........................................................................................................3

1.1. About UNICORN...............................................................................................4

1.2. About this manual.............................................................................................8

1.3. About the UNICORN user documentation..........................................................10

1.4. Quick Start Guide............................................................................................13

2. UNICORN concepts............................................................................................................14

2.1. Concept definitions.........................................................................................15

3. How to start your system...................................................................................................18

3.1. How to start UNICORN and log on....................................................................19

4. How to create a method.....................................................................................................21

4.1. How to use the Method Wizard.........................................................................22

4.2. How to use text instructions.............................................................................27

5. How to perform method runs..............................................................................................29

5.1. How to run your method...................................................................................30

5.2. The System Control module..............................................................................33

5.3. How to change the way your run is displayed......................................................35

5.4. Manual control................................................................................................37

6. Scouting...........................................................................................................................39

6.1. How to set up a Scouting Scheme.....................................................................40

7. How to view results...........................................................................................................42

7.1. How to view and edit your result layout..............................................................43

7.2. How to view the run documentation..................................................................46

7.3. How to compare curves....................................................................................47

8. How to print your chromatograms.......................................................................................49

8.1. How to print active chromatograms...................................................................50

9. How to evaluate and save the results.................................................................................52

9.1. How to perform a basic peak integration............................................................53

9.2. How to save the results....................................................................................56

10. How to create and print reports........................................................................................57

10.1. How to print an existing report format.............................................................58

10.2. How to edit an existing report format...............................................................59

10.3. How to create and print a customized report format..........................................61

Table of Contents

• p i

Table of Contents

03-0014-92 • p ii

Introducing UNICORN1

This chapter contains:

• An overview of the UNICORN™ system and general information that you needbefore you can start.

• Information about the user documentation for UNICORN and how to use it.

• A Quick Start Guide that can be used as a shortcut for experienced users thatwant to start right away.

About this chapter

This chapter contains these sections.In this chapter

SeeTopic

1.1About UNICORN

1.2About this manual

1.3About the UNICORN user documentation

1.4Quick Start Guide

Introducing UNICORN 1

• p 3

About UNICORN1.1

This section is a general overview of the UNICORN system.Introduction

UNICORN is a complete package for control and supervision of chromatographysystems. It consists of control software and a controller card for interfacing thecontrolling PC to the chromatography liquid handling module.

What isUNICORN?

Liquid chromatography is used in separation processes, for analytical purposes orin the biochemical process industry.

UNICORN runs on a PC under Microsoft® Windows® 2000 or MicrosoftWindows XP. It is designed to run under English keyboard settings.

Operating environ-ment

Note: Microsoft and Windows are registered trademarks of the MicrosoftCorporation in the United States and/or other countries.

Most Windows functions are also available in UNICORN, including:

• cut and paste

• right-click short-cut menus

Note: Drag and drop is not available. File and folder handling in UNICORN alsodiffers from the general Windows file manager standard.

Windows func-tions

UNICORN can be used with a number of systems including

• ÄKTA™ design systems

• BioProcess™ systems

Note: All examples in this guide are based on an ÄKTAexplorer™ 100 system thatoperates with the E100F400 strategy. If you use another system you may find thatthe descriptions and instructions do not match your system on every point. In thatcase you also need to refer to the user documentation for your specificchromatography system.

Compatible chro-matography sys-tems

1 Introducing UNICORN1.1 About UNICORN

03-0014-92 • p 4

UNICORN can be installed on a stand-alone computer to control only a single,locally attached system. However, a stand-alone computer can control up to amaximum of four separate systems. In a network installation each computerworkstation can operate many systems regardless if they are locally connected ornot. Each system can only be operated by one workstation at a time, but severalmay view the output data.

System networks

The UNICORN control software consists of four integrated modules:

FunctionModule

Software modules

File handling and administration, e.g.definition of systems and user profilesetc.

UNICORN Manager

To create and edit methods for pre-programmed control of chromato-graphy systems.

Method Editor

To control and monitor the separationprocesses online, through method-based or manual control.

System Control

To evaluate and present stored resultsfrom separation processes.

Evaluation

Note: All modules are active when the program is operating, and are not closedwhen they are minimized. A minimized System Control unit may control a process.


• p 5

The work flow in UNICORN can be divided into four distinct stages. Each stageis described in separate chapters in this manual. The flow chart below shows thework flow stages.

Work flow

An online help utility is included in the UNICORN software. The table belowdescribes how to access the help utility.

Then...If you want to access...

Help functions

open the Help menu in any of the software modules.the general help utility.

• click the Help button in the dialog box.

or

• press the F1 key on your keyboard.

context-specific helptopics.

The table below describes the main security functions in UNICORN:

FunctionFeature

Security

Only authorized users can accessUNICORN. Each user is assigned anaccess level, which defines the func-tions that the user is permitted to use.

Access Security

A running system can only be con-trolled from one connection. Systemsmay be locked with a password toprevent other, un-authorized usersfrom changing parameters.

Connection Security

1 Introducing UNICORN1.1 About UNICORN

03-0014-92 • p 6

FunctionFeature

Result files from an ongoing separa-tion run can be saved automatically atpreset intervals to minimize data lossif the system fails. The results aresaved locally if the network communic-ation fails.

Data Security

Method and result files can be signedelectronically for enhanced securityand accountability.

Electronic Signatures


• p 7

About this manual1.2

This section provides a general description of the manual, the contents and thepre-requisites for the examples and instructions that are presented in the GettingStarted guide.

Introduction

The purpose of Getting Started is to present a quick and easy guide to the systemfor a user with limited or no experience of UNICORN. The work flow is presentedin the form of practical instructions for how to operate a model system. Theseinstructions form a basic framework that the reader can expand on by readingselected parts in the other manuals.

The purpose ofGetting Started

UNICORN software can be used in numerous possible system variations. Forpractical reasons the user documentation is based on a model system that consistsof:

The model system

• ÄKTAexplorer 100

• Strategy E100F400

• Frac-950

Note: If you use another system you may find that the descriptions and instructionsdo not match your system on every point. In that case you also need to refer to theuser documentation for your specific chromatography system.

The examples and instructions that are presented in this manual are deliberatelylimited to a minimum to reduce the number of pages that you will need to read.In almost all cases you will find additional information about each topic in theUser Reference Manual.

Refer to othermanuals

The manual is divided into chapters. Each chapter starts with a brief overview thatpresents the contents and the headings for the sections that the chapter contains.The section begins with an introduction that summarizes the content.

Document struc-ture

A section is divided into blocks of information with separating lines. The blocksare identified by labels in the margins. This makes it easier for you to quickly scana page to find the exact topic you are looking for.

Getting Started contains ten chapters that can be sorted logically into 4 majortopics:

The manual con-tents

ChapterTopic

1. Introducing UNICORN

2. UNICORN concepts

Background information.

1 Introducing UNICORN1.2 About this manual

03-0014-92 • p 8

ChapterTopic

3. How to start your system

4. How to create a method

How to prepare the system.

5. How to perform method runs

6. Scouting

How to run the system.

7. How to view results

8. How to print chromatograms

9. How to evaluate and save the res-ults

10. How to create and print reports

How to use the results.

Menu commands, field names and other text items from the software are quotedexactly as they appear on the screen, in a bold typeface:

• Example: Run Setup

Search paths are shown in a bold typeface with a separating colon between eachlevel:

• Example: View:Windows:Customise (i.e. the menu command Customise in thesub-menu Windows from the View-menu).

Text entries that UNICORN generates or that the user must type are representedby a monotype typeface:

• Example: Connection change

Typographicalrepresentations

The following pre-requisites must be fulfilled before you can use this manual theway it was intended:

• You need to have a general understanding of how your PC and Windows works.In most cases universal computer functions will not be explained.

• UNICORN must be installed and configured correctly on your computer.

• You need to understand the concepts of liquid chromatography. Terminologyand functionalities will be explained only when they differ from normal practise.

• Before you try to operate a chromatography system based on the instructionsin this manual you need to study and understand the safety information that ispart of the system documentation.

Pre-requisites


• p 9

About the UNICORN user documentation1.3

The user documentation for UNICORN is divided into three separate manuals.This section is an overview of the contents and the relationship between themanuals.

Introduction

The three manuals are:

• Getting Started with UNICORN(See section 1.2 About this manual on page 8)

• UNICORN User Reference Manual

• UNICORN Administration and Technical Manual

The manuals

The questions and answers in the table below describe the features of the GettingStarted manual.

AnswerQuestion

User info aboutGetting Started

Users that are new to the UNICORNsystem and with limited experiencefrom other chromatography systems.

Who should read Getting Started?

A basic knowledge of PC andWindows functions and an understand-ing of the concepts and terminologyof liquid chromatography.

What do I need before I start?

Basic descriptions of UNICORN andits use, based on a model system.

What are the contents of GettingStarted?

Read in front of your computer andtest the instructions at the same time.

How should I use Getting Started?

When you need:

• more in-depth information,

• alternative ways to perform a task.

When do I need to refer to the UserReference Manual?

When you need:

• general information about the net-work functions of UNICORN,

• to set up and configure aUNICORN workstation in a net-work,

• to trouble-shoot a workstation withnetwork problems.

When do I need to refer to the Admin-istration and Technical Manual?

1 Introducing UNICORN1.3 About the UNICORN user documentation

03-0014-92 • p 10

The questions and answers in the table below describes the features of the UserReference Manual.

AnswerQuestion

User info aboutthe User ReferenceManual

• Users that are experienced withprevious UNICORN system ver-sions.

• Users with vast experience fromother chromatography systems.

Who should read the User ReferenceManual?

• Knowledge of PC and Windowsfunctions.

• An understanding of the conceptsand terminology of liquid chroma-tography.

• Preferably some previous experi-ence with UNICORN.


• Detailed descriptions ofUNICORN.

• Instructions on how to use the sys-tem, with suggested alternatives.

Note: Most instructions are based ona model system.

What are the contents of the UserReference Manual?

Depending on your previous experi-ence you can either read

• whole chapters from the beginningto the end

• only selected sections for reference.

How should I use the User ReferenceManual?

When you need brief, step-by-step in-structions for a selected task.

When do I need to refer to GettingStarted?

When you need

• general information about the net-work functions of UNICORN.

• to set up and configure aUNICORN workstation in a net-work.

• to trouble-shoot a workstation withnetwork problems.

When do I need to refer to the Admin-istration and Technical Manual?


• p 11

The questions and answers in the table below describes the features of theAdministration and Technical Manual.

AnswerQuestion

User info aboutThe Administra-tion and TechnicalManual

System administrators.Who should read the Administrationand Technical Manual?

• General knowledge of UNICORN.

• Knowledge of PC, Windows andgeneral network administrationfunctions.

• An understanding of the conceptsand terminology of liquid chroma-tography.


Detailed instructions of

• how to install and maintainUNICORN in a network environ-ment.

• how to create and administrate userprofiles.

Note: Most instructions are based ona model system.

What are the contents of the Adminis-tration and Technical Manual?

• If you are an experienced adminis-trator of previous UNICORN ver-sions you can read selected sectionsfor reference.

• If this is your first experience ofUNICORN administration we re-commend that you study themanual in detail.

How should I use the Administrationand Technical Manual?

When you need brief, step-by-step in-structions for a selected task.

When do I need to refer to GettingStarted?

When you need more informationabout the basic functions ofUNICORN.

When do I need to refer to the UserReference Manual?

1 Introducing UNICORN1.3 About the UNICORN user documentation

03-0014-92 • p 12

Quick Start Guide1.4

This guide is intended for users who are fully familiar with the safety precautionsand operating instructions that are described in all manuals, i.e. experienced usersof previous versions of UNICORN. The instructions assume that all installationswere made according to the instructions, that the model system is used and isconnected.

Introduction

The table below describes the easiest way to create a method, run the system andgenerate a printed chromatogram. The procedure is based on an Instant run.

ActionStep

Quick Start in-structions

Click the Instant run icon in the UNICORN Manager.

Result: The Instant run dialog box opens.

1

• Select Wizard.

• Select a system (if necessary).

• Click the Run button.

Result: The Method Wizard opens in the System Control module.

2

Go through all selections on the Method Wizard pages. Click the Nextbutton to proceed from page to page.

3

Click the Run button on the last page.

Result: The start protocol opens.

4

Verify the method on the Variables page and change values as re-quired. Click the Next button to proceed through several pages.

5

Select Print_Chromatogram in the Evaluation procedures page.

Result: A printout will automatically be generated after the run.

6

Click the Start button on the last page.

Result: The run starts.

7


• p 13

UNICORN concepts2

This chapter contains definitions and descriptions of some of the specific conceptsthat are presented in this manual and in the other UNICORN manuals. Generalconcepts and common chromatography terminology are not explained here.

Introduction

This chapter contains this section.In this chapter

SeeTopic

2.1Concept definitions

2 UNICORN concepts

03-0014-92 • p 14

Concept definitions2.1

• This section contains explanations and definitions of a number of UNICORNconcepts that are used in this manual.

• The concepts are sorted in alphabetical order.

Note: The section also lists some concepts that are described only in the UserReference Manual. These concepts are included for reference since they may befound in menus and dialogs that you will use while working with this manual.

Introduction

Systems settings or method instructions specify acceptable limits for monitor signalsduring a separation run. An Alarm dialog box will be displayed on the screen andan optional alarm can sound if a specified limit is exceeded. The system will bepaused.

Alarms

Note: Refer to the User Reference Manual for information about Alarms. The topicis not covered in this manual.

You can perform a Batch run of a number of result files in the Evaluation module.The files do not have to be open and the run operates in the background. Theprocedure is useful if you want to print a number of results with the same settings,or if you want to perform integration with the same parameter settings on manyresults.

Batch run

Note: Refer to the User Reference Manual for information about Batch runs. Thetopic is not covered in this manual.

BufferPrep is a function to prepare a buffer of different pH and salt concentrationsonline from four stock solutions. This eliminates the need to manually preparenew buffers every time the pH needs to be changed.

BufferPrep

Note: BufferPrep is only available for some ÄKTAdesign systems. Refer to the UserReference Manual for information about BufferPrep. The topic is not covered inthis manual.

A chromatogram is a collection of data represented by a number of curves thathave been created during a separation run, including UV, conductivity, pH, fractionmarks etc. The original raw data curves cannot be deleted or modified. They canbe used as a basis for evaluation procedures and subsequent creation of new curves.

Chromatogram

A chromatogram can also contain curves that have been created and saved duringan evaluation session.

The monitor signals from the chromatography run are displayed graphically ascurves.

Curves

UNICORN concepts 2

• p 15

The program instructions for a chromatography run are defined in a Method. AMethod is normally divided into blocks that represent steps in the separation process.Each block consists of a series of instructions that request specific operations inthe system.

Method

MethodQueues are used to link several methods together, on the same or on differentsystems.

MethodQueue

Example: A MethodQueue can be set up to conduct a CIP study of a number ofcolumns, through a controlled series of scouting runs.

Note: A method can be placed in a MethodQueue if the system is busy when theoperator wants to run the method. Refer to the User Reference Manual forinformation about MethodQueues. The topic is not covered in this manual.

The Method Wizard is a user-friendly tool to create new methods. The Wizard takesthe user step-by-step through the creation process.

Method Wizard

Method Wizards are supplied with UNICORN installations for ÄKTAdesign systems.

UNICORN creates Result files when a method is run. The Result files contain:

• Run data from the monitors in the chromatography system

Example: UV absorbance, flow rate, conductivity, etc.

• Documentation from the run

Example: Logbook entries, calibration settings, scouting parameters, text methodetc.

• Saved results from evaluations of the run data

Example: Peak integrations, simulated peak fractionations etc.

Result files

Scouting is used to repeat a series of Method runs automatically with predeterminedchanges in the values for one or more Variables. A Scouting Scheme is defined aspart of the method.

Scouting

Scouting is used for optimizing chromatographic processes.

2 UNICORN concepts2.1 Concept definitions

03-0014-92 • p 16

Part of the UNICORN software is specific for the system that it is set up to operate.The system specific part is usually referred to as the Strategy. The Strategy definesavailable method and manual instructions, system settings, run data, curves andmethod templates.

Strategy

Note: The examples in this guide are generally based on the E100F400 strategy.

Templates are basic methods that can be used as a starting point for developingcustomized methods. The method variables in a suitable Template is adjusted tocreate a method for another application.

Template

Values at breakpoints in the Method and instruction parameters may be defined asVariables. Variables makes it easy to adapt a method to a particular chromatographyrun.

• A framework Method with default parameters can be changed to create variants.

• A Method can be used in automatic Method Scouting, where one or moreparameter Variables are changed systematically.

Variable

Systems settings or method instructions specify acceptable limits for monitor signalsduring a separation run. A Warning dialog box may be displayed on the screen ifa specified limit is exceeded. The system will still continue to run after a Warning.

Warnings

Note: Refer to the User Reference Manual for information about Warnings. Thetopic is not covered in this manual.

UNICORN concepts 2

• p 17

How to start your system3

The first step when you begin your work with UNICORN is to start the system.This chapter describes how to start the program and log on as a user.

Introduction

This chapter contains the section below.In this chapter

SeeTopic

3.1How to start UNICORN and log on

3 How to start your system

03-0014-92 • p 18

How to start UNICORN and log on3.1

This section describes how to start the UNICORN program and how to log on asa user.

Introduction

The system administrator creates and defines users and may also create your initialpassword. (You may change the initial password to another later). The programcan also be set up so you can logon without a password.

Username andpassword

Note: If UNICORN is already started by a previous user, proceed to "How to logon".

There are two ways to start the program:

then...If you start with...

How to start theprogram

double-click the icon.a UNICORN icon onyour desktop,

locate the program under Programs:Unicorn and clickUNICORN.

the Windows Startmenu,

This is an illustration of the UNICORN Logon dialog box:The Logon dialogbox

Note: The Logon dialog has a Password text box only if a password is required.

How to start your system 3

• p 19

The table below describes how to log on to UNICORN.

ActionStep

How to log on

• Select Tools:Logon.

or

• Click the Logon/Logoff icon in the UNICORN Manager.

1

Select your username in the list.2

Type your password (optional).3

Click OK.4

The program has four windows or modules. When you start the program and logon you work in the UNICORN Manager. UNICORN also automatically opens theMethod Editor, the System Control and the Evaluation modules. These modules areminimized until you activate them.

The four programwindows

Always log off when you leave the computer to prevent others from accidentallychanging or deleting your files, or disturbing your method runs. There are twoways to log off:

• Select Tools:Logoff.

or

• Click the Logon/Logoff icon.

Log off after youare finished

3 How to start your system3.1 How to start UNICORN and log on

03-0014-92 • p 20

How to create a method4

Chromatography runs are programmed as Methods in UNICORN. Before you canproceed with a chromatography run you need either to use an existing method orcreate a new method. This chapter describes two different ways to create newmethods.

Introduction

This chapter contains the sections below.In this chapter

SeeTopic

4.1How to use the Method Wizard

4.2How to use text instructions

How to create a method 4

• p 21

How to use the Method Wizard4.1

This section describes how to use the Method Wizard to create a new method. Formost purposes, customized methods can be created simply by setting appropriatevalues for the method variables.

Introduction

Wizards are available for ÄKTAdesign systems delivered with standard strategies.If your system is a process systems please refer to the system documentation.

Are wizards al-ways available?

The Method Wizard consists of a number of dialog boxes in which you answerquestions and receive instructions on how to create your method. The options insubsequent boxes depend on the choices you have made in the initial dialog box.

How does thewizard work?

The table below describes an example of how to create a method with the MethodWizard.

ActionStep

How to create anew method

Click the Method Wizard icon in the Method Editor module.

Result: The Method Wizard dialog box appears.

Note: If several systems are available you must first select whichsystem you want to use.

1

4 How to create a method4.1 How to use the Method Wizard

03-0014-92 • p 22

ActionStep

Select the appropriate parameter values and click Next.

Note: Click Set Default to restore all settings to the default values.

2

On each new dialog page, select the appropriate parameter valuesand click Next to continue.

3

Click Finish in the last window.

Result: Run Setup opens.

4


• p 23

The Run Setup consists of a number of tab pages. Click the tabs at the top of thedialog box to select a tab page.

The Run Setup

The method is represented by a number of blocks on the Variables tab. The blocksare typical steps in a chromatographic run.

Run Setup Vari-ables

Each block contains a number of Variables, with suitable default values that canbe changed to suit your application. Some values are normally hidden. Click thecheckbox Show Details to display hidden variables.

When you select a column, default values will be set for:

• The correct column volume.

• The recommended flow rate.

• The correct pressure limit.

Note: If you exceed the recommended values for the selected column you willreceive a warning when you save your method.

Default values


03-0014-92 • p 24

The Gradient tab shows the method graphically:The Gradient page

The length of each block is marked at the bottom of the graph.

• Click the x-axis to view the method in time, volume or column volumes (CV).

The Start Protocol tab shows the items that will be displayed as dialog boxes in theRun Setup sequence.

• Click the checkboxes for the items that you want to select.

The Start Protocol


• p 25

Before you can run your method you must save it.

• Select File:Save.

or

• Click the Save icon.

How to save yourmethod


03-0014-92 • p 26

How to use text instructions4.2

Sometimes you need more advanced editing facilities, which are available whenyou work directly in the Text Instructions Editor in the Method Editor. This sectionis a brief description of this process.

Introduction

You can use the Text Instructions in the Method Editor to build your method stepby step. You can also use the editor to modify instructions in methods created bywizards or based on templates.

The Text Instruc-tions Editor

Use text instructions when you want to:

• change selected instructions in the method, e.g. the outlet valve position,

• add blocks or instructions, e.g. Watch instructions,

• change method instructions to adapt to non-standard system configurations,

• create new methods for applications not covered by the supplied templates orwizards.

When do I useText Instructions?

The system strategy determines the available instructions. A method that isdeveloped for one system may not be valid for another.

Valid instructions

The table below describes how to open the Text Instructions Editor.

ActionStep

How to open theEditor

Select the Method Editor module.1

Select File:New.

Result: The New Method dialog opens.

2

• Select a system (if more than one is available)

• Select Method Editor and click OK.

Result: The Method Editor module opens in text edit mode.

3

Proceed with step 5 and 6 if the screen is blank.4

Click the Customise Panes icon.5


• p 27

ActionStep

Select Text and Instruction Box and click OK.6

The text instructions are selected in the Instruction box in the lower part of theMethod Editor. Applicable parameters can be edited for each selection. Use thebuttons to Insert, Change, Replace or Delete the selected instructions. All text entriesare shown in the Text pane.

The illustration below shows the Instruction box:

How to enter TextInstructions

4 How to create a method4.2 How to use text instructions

03-0014-92 • p 28

How to perform method runs5

Once you have a defined method in place you can perform a method run. You canmonitor its progress in the System Control module. This chapter describes how toperform the method run, the data display, how you can focus on the informationyou are interested in and make adjustments while the method run is in progress.

Introduction


SeeTopic

5.1How to run your method

5.2The System Control module

5.3How to change the way your run is displayed

5.4Manual control

How to perform method runs 5

• p 29

How to run your method5.1

Follow the instructions in this section if you want to start a separation run basedon the method you created and saved in the previous chapter. Also use thisprocedure if you want to run other methods.

Introduction

Before you can start a run, you must connect to the system. Open the System Controlwindow and look at the Connection panel in the Run Data section. If you are notconnected the panel will show the text NO. Once you are connected the text changesto YES. Refer to the User Reference Manual if your system is not connected.

Connect to thesystem

After the system is connected it must be prepared. Verify that this already has beendone or refer to your system documentation for the correct procedure.

Prepare the system

You initiate the method run in a series of dialog boxes in the Run Setup in theSystem Control module. The Start Protocol for the method decides which pages youneed to fill in. The steps in the table below is an example of a Run Setup sequence.When you are finished in one dialog box you click Next to proceed.

How to start amethod run

ActionStep

• Select the File:Run menu command

or

• click the Run toolbar icon.

1

The Run Setup opens. Select a method and click OK.2

If your method run involves a fraction collector, i.e. Frac-950, thefirst step is usually the setup dialog box. See "How to set up Frac-950" below.

3

The Variables dialog box opens next. Verify and fine tune yourmethod before you proceed.

4

In the Notes dialog box some information can already be present.Click the Start Notes tab and add your own comments.

5

5 How to perform method runs5.1 How to run your method

03-0014-92 • p 30

ActionStep

In the Questions dialog box some questions may be mandatory andmust be answered before you can start.

6

In Evaluation Procedures you select the automated operations youwant the system to perform after the UNICORN run. SelectPrint_Chromatogram for an automatic print-out after the run.

7

The Method Information page is a summary of information about therun. Click Next to proceed to name your result file and define whereit should be stored.

8

In the Frac-950 setup dialog box you can define the order of fractionation and setup the last tube. The system will be paused when the last tube is reached and thefractionation will stop.

The illustration below shows the Frac-950 setup dialog box:

How to set upFrac-950

The final step before starting is to name your result file and define where it shouldbe stored. The default file name is defined in the Result Name page of the Run Setup.By default the result file name will either be the same name as the method, the dateof the run or a pre-defined name. The name is followed by a three-digit sequencenumber starting with 001. You can change this name and select a new directoryby clicking Browse.

How to name theresult file

You can also select to have the software add a unique identifier to the file name.


• p 31

Click the START button in the Result Name dialog box. This will initiate the methodrun and you can follow its progress in the System Control module.

How to start themethod run

5 How to perform method runs5.1 How to run your method

03-0014-92 • p 32

The System Control module5.2

All data on your separation run are displayed in the System Control module. Youhave a choice of four different panes that can be open one at a time or all at oncein separate parts of the window.

Introduction

The table below describes how to select the panes that are displayed in the SystemControl module.

ActionStep

How to select dis-plays

• Select the View:Panes menu command.

or

• Click the Customise Panes icon.

1

Click the check boxes for the panes that you want to display.2

Click OK.3

The Run Data pane shows the current values for the running parameters. When thesystem is running, the text Run is displayed in the Run Status panel. If the systemis operated manually the text Manual is displayed. The illustration below showsonly part of the Run Data pane.

The Run Datapane


• p 33

The Curve Data pane shows selected monitor signals as curves during your methodrun. All curves are stored in the result file. Normally the curves are scaled withauto scaling, i.e. the scale is adjusted continually to the highest and lowest valuesfor each curve.

The Curve Datapane

The Flow Scheme pane shows a schematic view of your system configuration.The Flow Schemepane

The Logbook pane is shown at the bottom. The Logbook shows exactly when theinstructions in the method were executed during the run. It also shows all manualinstructions that were performed and all alarms and warnings that were registered.

The Logbookpane

then....If you want....How to view asingle pane

right-click and select Maximize.to enlarge a pane

right-click and select Restore.to return to the origin-al size

5 How to perform method runs5.2 The System Control module

03-0014-92 • p 34

How to change the way your run is displayed5.3

There are a number of ways to change the way your method run data is displayedin the System Control module. This section describes a few of the options.

Introduction

The table below describes how to open the Properties dialog box to edit the waythe panes are displayed.

ActionStep

How to edit thepane displays

Right-click in the pane you want to edit.1

Select Properties from the menu.2

Click the tab for the pane you want to edit.

Result: The page shows all the options that you can select in thisparticular pane. You also have a choice of different styles, colorsand axis layouts for your curves.

3

You can select which curve the Y-axis scale refers to in two different ways:

• Click the curve name at the top of the Curve Data pane.

or

• Click the Y-axis scale to toggle between the curve scales.

How to changethe Y-axis scaledisplay

The table below describes how to set a fixed value range for the Y-axis for a selectedcurve in the Properties dialog box.

ActionStep

How to set fixedvalues for the Y-axis

Click the Y-axis tab.1


• p 35

ActionStep

Click the curve you want to edit.2

Click the Fixed radio button.3

Type a minimum and a maximum value. The maximum range valuesallowed are shown above the entry boxes.

4

Repeat steps 2 to 4 for all other curves you want to edit.5

Click OK.6

5 How to perform method runs5.3 How to change the way your run is displayed

03-0014-92 • p 36

Manual control5.4

In some applications you may want to change some parameters manually duringa run. This section exemplifies how you can change the pump flow manually.

Introduction

The table below describes how to change the pump flow.

ActionStep

How to changethe pump flow

Select Manual:Pump.

Result: the System Pump Instructions dialog box opens The Pumpradio button is selected.

1

Click the function you want to change, i.e. Flow.2

Enter a new value (FlowRate) under Parameters. You can use the ar-rows to step the value up or down.

3

• Click Execute to execute the instruction immediately.

or

• Click Insert to add the instruction to the list below the instructionsmenu.

Note: If there are instructions on the list, the Execute button willexecute all instructions on the list at the same time.

4

Click Close to close the dialog box.5

The illustration below shows the System Pump Instructions dialog box:The System PumpInstructions box

Note: The parameter values will be updated continually during the run if the Autoupdate checkbox is selected.


• p 37

Click the End button to end the method run before it is finished. You can save thepartial result the same way that you save a completed run.

How to end yourrun manually

5 How to perform method runs5.4 Manual control

03-0014-92 • p 38

Scouting6

You can use Scouting to automatically repeat a series of method runs with asystematic variation of one or more parameters. This chapter gives an example ofhow to use Scouting.

Introduction


SeeTopic

6.1How to set up a Scouting Scheme

Scouting 6

• p 39

How to set up a Scouting Scheme6.1

When you prepare your method you can define a number of variables in the methodfor Scouting. The definition of how the series of method runs will be performed iscalled a Scouting Scheme.

Introduction

You can use Scouting for example:

• to screen for the best column,

• to find the optimal pH,

• to test column capacity (sample volume),

• to find the optimal flow rate for binding and elution,

• to optimize gradient length and slope.

When to useScouting

Scouting Schemes are defined in the Run Setup in the Method Editor. Before you canstart you need to create a new method, for example by using a method wizard.You can also add a Scouting Scheme to an existing method.

Before you start

Note: Variables must be defined for Scouting.

The table below describes how to set up a Scouting Scheme.

ActionStep

Example: Flow-Rate Scouting

Click the Scouting tab in the Run Setup.

Result: A Scouting Variables dialog box opens.

1

• Click the checkbox for the variable Flow_Rate and any othervariable you wish to scout, e.g. BufferPrep_pH.

• Click OK.

Result: The selected scouting variables will appear in a column.

2

Click Add to create a second column for the next run. This columnwill have the same values as the previous column.

3

Change variable values in the same manner as on the Variables page.

See 4.1 How to use the Method Wizard on page 24

4

Repeat steps 3 and 4 until you have defined all the runs you require.6

Select the File:Save menu command.7

6 Scouting6.1 How to set up a Scouting Scheme

03-0014-92 • p 40

The illustration below shows the Scouting tab in the Run Setup.The Scouting tab

The table below describes how to edit a Scouting Scheme:

then...If you want...

How to edit aScouting Scheme

click the column header.to select a run

select the column and click the Delete button.

Result: The column is removed.

to delete a run

right-click the column header.

Result: The column is marked Excluded and will notbe executed. It will still remain in the scouting scheme.

to exclude a run

select the column to the left of the position where youwant to insert the new column, and click the Insertbutton.

Result: A new column is added immediately to theright of the selected column.

to insert a new run

Click the Define button.

Result: The Scouting Variables dialog box opens.

to define new scoutingvariables

When the method is started the scouting runs are performed automatically. Eachmethod run in the Scouting Scheme generates a separate result file, which is storedin a special scouting directory.

The scouting res-ults

Scouting 6

• p 41

How to view results7

This chapter describes how to view the results from your method run in theEvaluation module.

Introduction


SeeTopic

7.1How to view and edit a result layout

7.2How to view the run documentation

7.3How to compare curves

7 How to view results

03-0014-92 • p 42

How to view and edit your result layout7.1

This section describes the basics of how to view and edit the layout of your resultsin the Evaluation module.

Introduction

The table below describes how to view the results.

ActionStep

How to open theresult file

Complete the method run.1

Locate the result file in the Results pane in the UNICORN Managermodule.

2

Double-click the file.

Result: The file opens in the Evaluation module.

3

The result file is opened in a Chromatogram window. The default view shows allthe curves as in the illustration below.

The Raw Chroma-togram Data

The table below describes how to change the layout of the Chromatogram display.

ActionStep

How to changethe chromatogramlayout

Right-click in the Chromatogram window and select Properties.

Result: The Chromatogram Layout dialog box opens.

1

Click the Curve tab.2

Click the check-boxes to de-select the curves that you do not wantto display.

3

How to view results 7

• p 43

ActionStep

Click OK.4

Your selections determine the curves that are displayed in the Chromatogram windowand shown on printouts.

Raw data curvesare saved

Note: The original raw data curves can never be modified, renamed or deletedfrom the result file.

Normally the curves are auto scaled, i.e. the highest and lowest values for eachcurve set the scale. The table below describes how to change auto scaled Y-axisscales to fixed values in the Chromatogram Layout dialog box.

ActionStep

How to changethe Y-axis scale

Click the Y-axis tab.1

Click the curve you want to edit.2

Click the Fixed radio button.3

Enter new minimum and maximum values.4

Repeat steps 2 to 4 for all other curves you want to edit.5

Click OK to execute the changes and close the window.6

You can select which curve the Y-axis scale refers to in two different ways:

• Click the curve name at the top of the Curve Data window.

or

• Click the Y-axis scale to toggle between the curve scales.

How to changethe Y-axis scaledisplay

The table below describes how you can zoom in on a curve to view more details.

ActionStep

How to viewcurve details

Place your cursor in a corner of the area you want to enlarge.1

Press and hold the left mouse button and drag diagonally over thearea.

Result: A rectangle appears.

2

Release the mouse button.

Result: The area within the rectangle is enlarged.

3

7 How to view results7.1 How to view and edit your result layout

03-0014-92 • p 44

ActionStep

Repeat steps 1 - 3 to enlarge the selected area further.4

Right-click and select Undo Zoom to zoom out one step.5

Right-click and select Reset Zoom to return to the full window view.6

You can save your edited layout. It can be applied later to any result file. Followthe steps in the table below.

ActionStep

How to save alayout

Right-click and select Properties to open the Chromatogram layoutdialog box.

1

Select the Layout library tab.2

Click the Save current layout as... button.3

Enter a name for the layout and click OK.4

Select a layout from the Saved layouts list in the Layout library. Click the Applyselected layout button.

How to apply asaved layout


• p 45

How to view the run documentation7.2

The full documentation of a method run is stored in the result file. You can viewthis information in the Documentation dialog box in the Evaluation module. Thissection gives an example of how to view the Logbook.

Introduction

Maximize the Evaluation module and either:

• Select the View:Documentation menu command

or

• Click the View Documentation icon.

How to open therun documenta-tion

This is an illustration of the Documentation dialog box with the Logbook tab selected.Click the tabs to view other information.

The documenta-tion window

7 How to view results7.2 How to view the run documentation

03-0014-92 • p 46

How to compare curves7.3

You can import or copy curves from different method runs into one chromatogramfor comparison. This section is an example of how you can use the function Opento compare to import curves.

Introduction

First open the dialog box Open Curves to Compare. You can either:

• Select File:Open to compare:Curves.

or

• Click the Open curves to compare icon.

Open the dialogbox

The illustration below shows the dialog box Open Curves to Compare.The Open curvesto compare dialogbox

The table below describes how to import curves and store them in a newchromatogram.

ActionStep

How to importthe curves

Define search criteria for the folder, result, chromatogram and/orcurve name. Use the Browse command buttons.

1


• p 47

ActionStep

Click Search.

Result: A list of found curves is displayed.

2

Repeat step 1 - 2 to add more curves to the list.3

• Select the individual curves that you want to import.

or

• Click Select all to import all curves.

4

Click the checkbox Store in new chromatogram and enter a name inthe text box (Compare is default).

Note: You can also save the curves in the active chromatogram.

5

Select one of the Curve options: Overlay, Stack or Mirror.

Result: This will decide how the curves are displayed.

6

Click OK.7

If you selected the Stack option the Shift Curves by Offset dialog box is openedautomatically. Adjust the offset distance between the curves to a suitable value.

How to set thestack offset

7 How to view results7.3 How to compare curves

03-0014-92 • p 48

How to print your chromatograms8

This chapter describes how to print open chromatograms. Proceed to 10 How tocreate and print reports on page 57 if you want to add text information to yourprints or create a complete report.

Introduction


SeeTopic

8.1How to print active chromatograms

How to print your chromatograms 8

• p 49

How to print active chromatograms8.1

This section describes how to print the chromatograms that are open in theEvaluation module.

Introduction

Open all chromatograms that you want to print in the Evaluation module beforeyou proceed.

Before you print

The table below describes how to print active chromatograms.

ActionStep

How to print

• Select the File:Print menu command.

or

• Click the Print icon.

Result: The Print Chromatograms dialog box opens.

1

Select print formats and layout options.2

Click Preview.

Result: The Customise Report window opens.

3

• Verify that the layout is correct.

• Click Edit Mode to make changes, e.g. change the order of thechromatograms. Click Preview to return to preview mode.

• Click Exit to return to the Print Chromatograms dialog box.

4

Click OK.5

8 How to print your chromatograms8.1 How to print active chromatograms

03-0014-92 • p 50

This is an illustration of the Print Chromatograms dialog box.

Note: The selected print format is outlined in red.

The Print Chroma-tograms dialogbox

How to print your chromatograms 8

• p 51

How to evaluate and save the results9

This chapter describes how to perform basic evaluation procedures and how tosave the results of the evaluations.

Introduction


SeeTopic

9.1How to perform a basic peak integration

9.2How to save the results

9 How to evaluate and save the results

03-0014-92 • p 52

How to perform a basic peak integration9.1

Peak integration is used to identify and measure curve characteristics, includingpeak areas, retention times and peak widths.

Introduction

A correct baseline must be calculated before the peak areas can be calculated. Thereare several alternative ways to perform this calculation:

• use automatic calculation,

• subtract a blank run curve from the source curve,

• use a Zero baseline, i.e. no baseline subtraction at all,

• re-use an existing baseline,

• edit a baseline manually from any curve in the chromatogram.

The default options are used for the example in this section, i.e. automaticcalculation (Calculate baseline) with a Morphological algorithm.

Baseline calcula-tions

The table below describes how to perform a basic peak integration.

ActionStep

How to performthe peak integra-tion

• Select the Integrate:Peak Integrate menu command.

or

• Click the Peak Integrate icon.

Result: The Integrate dialog box opens.

1

Select a source curve.2

Select a peak table destination from the Target peak table list.3

Select Calculate baseline in the Baseline list.4

Click OK.5

How to evaluate and save the results 9

• p 53

This is an illustration of the Integrate dialog box.The Integrate dia-log box

This is an illustration of the results after a peak integration.Peak integrationresults

Note: The peak table is displayed underneath the active chromatogram. The startpoint and end point of each peak are marked by vertical marks, drop-lines.

9 How to evaluate and save the results9.1 How to perform a basic peak integration

03-0014-92 • p 54

The peaks in the integrated chromatogram are automatically labelled with theirretention times. Several other peak characteristics are also calculated automatically.The table below describes how to display other characteristics.

ActionStep

How to displaypeak characterist-ics

Right-click in the active chromatogram.1

Select Properties.

Result: The Chromatogram Layout dialog box opens.

2

Click the Peak Table tab.3

Select the options that you want to display from the Select peaktable columns list.

Result: The selected items will be displayed in the peak table.

4

Click OK.5

This is an illustration of the Peak Table dialog box.The Peak Tabledialog box

How to evaluate and save the results 9

• p 55

How to save the results9.2

After you have finished the evaluation process you can save all the changes youhave made to the chromatograms, including newly created curves andchromatograms that you have imported and created.

Introduction

All the curves that you created during your manipulations will be saved in thechromatogram. Some of these curves may not be needed anymore.

• Select Edit:Delete:Curves to remove all unwanted curves.

Note: The original curves that were created during the run can never be deleted.

How to delete un-wanted curves

You can either save your results in the original file or in a new result file.

then...If you want to save...

How to save theresults

• choose File:Save

or

• click the Save icon.

in the original resultfile

• choose File:Save as.in a new result file

9 How to evaluate and save the results9.2 How to save the results

03-0014-92 • p 56

How to create and print reports10

The Evaluation module provides extensive tools to create detailed reports. Thischapter describes how to create reports.

Introduction


SeeTopic

10.1How to print an existing report format

10.2How to edit an existing report format

10.3How to create and print a customized report format

How to create and print reports 10

• p 57

How to print an existing report format10.1

This section describes how to use an existing report format to print a basic reportwith a chromatogram and text.

Introduction

The table below describes how to print a report.

ActionStep

How to print a re-port

• Choose File:Report.

or

• Click the Report icon.

Result: The Generate Report dialog box opens.

1

Select format (Global) Chromatogram.

Result: This creates a report containing the chromatogram and theanswers to the questions from the Run Setup Questions page.

2

Click Print.

Result: The Print dialog box opens.

3

Click OK.4

This is an illustration of the Generate Report dialog box.The Generate Re-port dialog box

10 How to create and print reports10.1 How to print an existing report format

03-0014-92 • p 58

How to edit an existing report format10.2

This section describes how to open an existing report format to edit the items thatare included in the report.

Introduction

The table below describes how to open and edit a report format.

ActionStep

How to open andedit a reportformat

• Choose File:Report

or



1

Select a report format.2

• Click Edit.

Result: The Customize Report window opens in edit mode.

3

• Double-click an item that you want to edit in the report.

Result: A Setup dialog box specific for the item opens.

4

• Edit the item.

• Repeat step 4 for all other items you want to edit.

5


• p 59

Once you have finished editing the report items you can apply the format.

then...If you want to...

How to apply thereport format

• click the Print icon.to print the report

• click the Preview button.to preview the changedreport layout

• choose File:Save As and enter a name for the reportformat.

to save the format

10 How to create and print reports10.2 How to edit an existing report format

03-0014-92 • p 60

How to create and print a customized report format10.3

You can select a number of different objects (including chromatograms, methods,images, free text etc.) and create a customized report format. The objects can bealigned, re-sized and positioned to fit your specific layout. This section describesonly some of the formatting options.

Introduction

The table below describes how to create a new customized report format.

ActionStep

How to create anew report format

• Choose File:Report.

or



1

Click the New button.

Result: The Create New Report Format dialog box opens.

2

Choose Customised Format and click OK.

Result: The Customise Report window opens.

3

Proceed to "How to add items to the empty report".4

The table below describes how to add items to your empty report format.

ActionStep

How to add itemsto the report

Select an information item.

• Click one of the item icons in the toolbar.

or

• Select an item from the Insert menu.

1


• p 61

ActionStep

Press and hold down the left mouse button, and drag out a box tothe size of the item you want to insert.

Note: The mouse pointer shows a symbol for the type of item youhave selected.

2

Release the mouse button.

Result: A Setup dialog box opens. The dialog box is specific to thetype of item that you want to insert.

3

Select the options you want.4

Click OK.5

Repeat steps 1 to 5 for each new item you want to insert.6

Click Preview to view the final results.7

The illustration below shows the Setup Chromatogram dialog box in the CustomiseReport window.

The Setup Chro-matogram dialogbox

The table below describes how to set up the page formatting.

ActionStep

How to set up thepage format

Double-click anywhere in the Customise Report window.

Result: The Page Setup dialog box opens.

1

Click the Page Setup tab and enter values for all Margins.2

Click the First Header tab and select the items that you want to in-clude in the first page header.

3

10 How to create and print reports10.3 How to create and print a customized report format

03-0014-92 • p 62

ActionStep

Click the Footer tab and select the items that you want to include inthe footer.

4

Click the Header tab and select the items that you want to includein the header.

Note: This tab is not available if you selected Same header on allpages in the Page Setup tab.

5

The illustration below shows the First Header tab in the Page Setup dialog box.The Page Setupdialog box


• p 63

03-0014-92 • p 64

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